In Situ Hybridization to Identify Stem Cells in the Freshwater Sponge Ephydatia fluviatilis

Chiaki Kojima; Noriko Funayama

doi:10.1007/978-1-0716-2172-1_17

In Situ Hybridization to Identify Stem Cells in the Freshwater Sponge Ephydatia fluviatilis

Methods Mol Biol. 2022:2450:335-346. doi: 10.1007/978-1-0716-2172-1_17.

Authors

Chiaki Kojima¹, Noriko Funayama²

Affiliations

¹ Department of Biophysics, Graduate School of Science, Kyoto University, Kitashirakawa-Oiwake, Sakyo-ku, Kyoto, Japan.
² Department of Biophysics, Graduate School of Science, Kyoto University, Kitashirakawa-Oiwake, Sakyo-ku, Kyoto, Japan. funayama.noriko.6a@kyoto-u.ac.jp.

Abstract

Sponges (Porifera) are a large phylum that includes an enormous number of species. They are classified into four classes. Among these four classes, class Demospongiae is the largest and contains more than 90% of sponge species. In the last decade, methodologies for molecular studies and sequencing resources in sponge biology have dramatically advanced and made it possible to clearly define particular types of cells based on the genes they are expressing. Here we describe in detail the method of high-resolution WISH (whole mount in situ hybridization) and dual color fluorescent detection of in situ hybridization (dual color FISH) that we have established to detect particular types of cells, especially their stem cells known as archeocytes, in juveniles of freshwater demosponge, E. fluviatilis.

Keywords: Archeocyte; FISH; Musashi; Piwi; Porifera; Stem cells; WISH.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Fresh Water
In Situ Hybridization
Porifera* / genetics
Stem Cells*